Bioresource Technology 193 (2015) 178–184

Contents lists available at ScienceDirec t

Bioresource Technology
journal homepage: www.elsevie r .com/locate/biortec h

Methods coagulation/flocculation and flocculation with ballast agent for

effective harvesting of microalgae
Kirill V. Gorin ⇑, Yana E. Sergeeva, Victor V. Butylin, Anastasiya V. Komova, Victor M. Pojidaev, Gulfiya U.
Badranova, Anna A. Shapovalova, Irina A. Konova, Pavel M. Gotovtsev
National Research Centre ‘‘Kurchatov Institute’’, NBICS-centre, Biotechnology and Bioenergy Laboratory, Moscow 123182, Russian Federation

highlights

Polyethylenoxide was successfully applied as flocculant for biomass sedimentation.

New method of biomass flocculation by combine using of polymer and FeCl3 is developed.
Application of flocculated biomass as ballast agent for microalgae harvesting.
a r t i c l ei n f o Chlorella vulgaris
Polymeric flocculants
Inorganic coagulants
Microalgae cultivation
Article history: abstract
Received 24 April 2015
Received in revised form 16 June 2015
The effects of coagulant (FeCl36H2O), various flocculants based on polyacrylamide (PAA), polyethylenoxide (PEO) and
Accepted 19 June 2015 Available
flocculated biomass as ballast agent, dosage and sedimental time on flocculation efficiency of harvesting Chlorella vulgaris
online 25 June 2015
GKV1 cultivated in a laboratory were investigated. The results of this work indicated that the flocculation efficiency achieved
about 90% after 5 min of sedimentation when adding of coagulant and flocculant mixture (FeCl3 50 mg/l + PEO based Sibfloc-
718 7.5 mg/l) or flocculant with ballast agent (Sibfloc-718 7.5 mg/l + 10% flocculated biomass). PAA and PEO showed good
flocculation efficiency at dosage of 0.025 and 0.015 g/l, respectively without pH adjustment. Finally, the most suitable
Keywords: flocculation method was discussed in this paper.
Flocculation
2015 Elsevier Ltd. All rights reserved.
1. Introduction production. The biomass concentration in a bioreactor depends on its type.
Thus, it may be up to 0.5 g/l in open reactors, and up to 5 g/l in closed systems
(Vandamme et al., 2013). Therefore, the requirement for microalgae recovery
With the growing world population and advance in the living standards of
is to somehow remove a significant amount of water (Schlesinger et al., 2012),
developing countries, the raw materials demand for food or feed uses is to
which definitely entails more power consumption when the microalgae are
increase by more than 50% in the next 20 years (Vandamme et al., 2013). Again,
conditioned for processing.
initiative efforts have been taken for transition from an economy based on fossil
fuels to a bioeconomy with renewable biomass to replace fossil oil as a source Using thermal or mechanical recovery techniques, such as induced drying,
of fuel for transport and a feedstock for chemical industry (Haveren et al., 2008). filter-pressing or centrifuging would need significant power consumption and
Microalgae are believed to be a new and more promising source of biomass for so impact the final product cost (Grima et al., 2003).
various applications (Vandamme et al., 2013). Efficiency of the flocculants for microalgal biomass sedimentation has been
Biofuel production using algal biomass calls a great interest in many shown by many studies on various phototrophic microbe cultures (Wan et al.,
research teams. Among the numerous species, phototrophic microalgae are 2015). There is evidence that with the use of commercially produced
certainly attractive, whose needs for growth are light, carbon dioxide and some flocculants, it would be possible to achieve 0.5 to 10–50 g/l of biomass
quantity of mineral substances. These are very important factors in biofuel concentration in the sediment (Wileman et al., 2011).

⇑
Corresponding author.
E-mail address: gkvbio@gmail.com (K.V. Gorin).

http://dx.doi.org/10.1016/j.biortech.2015.06.097 0960-8524/
2015 Elsevier Ltd. All rights reserved.
 K.V. Gorin et al. / Bioresource Technology 193 (2015) 178–184 179

The flocculant dose is also determined Study of the PAA flocculating properties for Scenedesmus sp. (0.54 g/l)
according to its type, and the algal species and found the following: for PAA concentration 0.05 g/l, flocculation efficiency

Coagulant or/and
flocculant

Culture medium
10%

90 %
Coagulant or/and
flocculant Flocculated biomass as
a ballast
Water to recycle

Flocculated biomass

Fig. 1. Block scheme of ballast agent application for microalgae harvesting.

concentration too. It may range from 10 to 50 was 60% in 10 min (Wan et al., 2015). Currently, research is also conducted
mg/l and more. Thus, processing 1000 l of on PAA modification to improve its flocculating properties (Banerjee et al.,
solution from a photobioreactor with biomass 2012).
concentration of 5 g/l may yield, in theory, up In addition to PAA, polyethylenoxide (PEO) is used as a flocculant in
to 5 kg of dry microalgae. For 50 mg/l dose, the treatment of wastewater (Tripathy and De, 2006). Most PEO-based
flocculation would have to use 50 g of commercial flocculants are weakly cationic, which makes them potentially
flocculant. Obviously, commercial production attractive for use as microalgae sedimentation agents.
would refer to the processing scale of dozens or Due to very low toxicity, PEO is widely used in pharmacy and biomedical
even hundreds of tons of culture broth, so that practice (Thompson et al., 2008), i.e., for immobilizing ferments (like
the flocculant costs would certainly contribute protease) (Vereschagin et al., 2001).
to the final product cost. Further, consider the
Chitosan is one of the first compounds of biological origin that was
flocculant types that can be found useful in
considered for use as a flocculant. This compound had been also considered
microalgae recovery from aqueous medium.
earlier as a flocculant for treating different sorts of wastewater. When it
The most common of these are compounds appeared, the microalgae harvest problem had once again excited interest in
based on aluminum and ferric salts, and this compound (Chang and Lee, 2012). The main difficulty with chitosan use
polymer-based flocculants. Metal salts were to harvest microalgae is that this compound is efficient at low pH values,
successfully used for microalgae sedimentation whereas the microalgal cultures are on the contrary characterized by pH above
(in particular, Dunaliella (Ben-Amotz and 7 (Chang and Lee, 2012). What should be also noted is poor solubility of
Avron, 1990), Microcystis sp. (El-Diwani et chitosan in water, which necessitates intensive stirring of the medium earlier
al., 2011)), but their use would lead to in floccule formation. The latter, in turn, would result in more power
undesired high concentrations of metals in the consumed for flocculation process as compared with traditional polymer-
sediment (Rwehumbiza et al., 2012). based flocculants.
Reportedly, magnesium hydroxide Yet, the above-discussed polymers (both synthesized, and chitosan) are
Mg(OH)2 and calcium hydroxide Ca(OH)2 can efficient no more in solutions of high ionic strength (Uduman et al., 2010), which
flocculate algae culture broth. Thus, the is something that hinders their use to harvest the algae growing in seawater.
sedimentation efficiency is shown in (Chen et
It should be mentioned, that solid–liquid separation through combined
al., 2013) to achieve 97.4% after 10 min of
coagulation and flocculation is frequently used at wastewater treatment stage:
settling Scenedesmus sp. with pH at 11.5
butchery wastewater (Aguilar et al., 2005) and also for sediment dewatering in
without flocculants added. Using sodium
pulp and paper, pharmaceutical, cosmetological, food and other industries (Lee
hydroxide with pH at 11 to flocculate
et al., 2011).
Botryococcus braunii was much more efficient
than the processing with aluminum sulfate and The very perspective approach to microalgae recovery is by using industrial
polysaccharide flocculant of bacterial origin coagulants and flocculants, agents that are used for fresh water (Harith et al.,
(Lee et al., 1998). 2009) or wastewater treatment (Jiang, 2015). These agents have different uses
according to their roles. Thus, the coagulant must destabilize the colloidal system
The commercial polymeric flocculants are
by neutralizing the forces of different origin that keep it stable. The flocculant’s
usually based on polyacrylamide (PAA)
role is to increase the size of flocs produced by coagulation and agglomerate the
(Bratby, 2006). Mainly, their applications
suspended particles for their removal (López-Maldonado et al., 2014). Currently,
include treatment of natural and wastewaters,
efforts are under way to speed up this process. One candidate decision is to add
and ore flotation tailings. Major consideration
suitable small-particle-size sand as a ballast agent. Sand particles would serve as
for wide use of PAA was high formation rate of
seed grains when flocculation proceeds, while accelerating the sedimentation due
a high molecular mass polymer. Other
to their weight (Jiménez et al., 1999). Using the recoverable ballastic agent can
considerations to be mentioned are low power
make the coagulation and sedimentation proceed faster. Clearly, the use of sand
consumption with the monomer separation as a
as ballastic agent in microalgae case would only complicate subsequent stages of
crystal, and that PAA has proved itself as
their processing. In view of this, the ballast sedimentation method for microalgal
nontoxic for humans, animals, fish and
biomass recovery calls for development of a new ballast agent, which have not
vegetation (Seybold, 1994).
been so far explored in microalgae application.
180 K.V. Gorin et al. / Bioresource Technology 193 (2015) 178–184

Fig. 2. Effect of the different dosages of four flocculants, (a) Superfloc N-300, (b) Superfloc A-100, (c) Superfloc C-492, (d) Sibfloc-718, at different sedimental time on the
It is known that bioflocculation is successfully 2. Methods
used for harvesting microalgae in facilities where
microalgae are used in wastewater treatment.
2.1. Microalgal culture and cultivation conditions
Some microalgal species flocculate more readily
than others and such naturally bioflocculating
The microalgae strain used in this study was C. vulgaris GKV1 (NRC
microalgae can be mixed with other species to
‘‘Kurchatov Institute’’ collection), which is widely accepted and used in various
induce flocculation. In many researches have been
science and technology applications.
shown that some bacteria and fungi have ability to
induce efficient bioflocculation of microalgae The microalgae C. vulgaris were grown on Basal medium (Yeh and Chang,
(Zhang 2012) consisted of (g/l): KNO3, 1.25; KH2PO4, 1.25; MgSO47H2O, 1; CaCl2,
flocculation efficiency of Chlorella vulgaris GKV1. 0.0835; H3BO3, 0.1142; FeSO47H2O,
0.0498; ZnSO47H2O, 0.0882; MnCl24H2O, 0.0144; MoO3, 0.0071; CuSO45H2O,
0.0157; Co(NO3)26H2O, 0.0049; EDTA2Na, 0.5. The culture medium prepared
and Hu, 2012; Surendhiran and Vijay, 2013; on filtered water, initial pH brought up to 7. Inoculum for experiments was
Talukder et al., 2014). Also it was proposed new cultivated in 250 ml Erlenmeyer flasks and then in 1000 ml and finally in 5000
effective method for harvesting of microalgae by ml flasks. The stirring was provided by the atmospheric air bubbling (250
bioflocculation of a non-flocculating microalga ml/min). The growth condition were kept constant under controlled temperature
with another autoflocculating microalga (Salim et (24 ± 1 C), with light intensity at 3000 lux at all times during the process. The
al., 2011). The advantage of this method in culture (20 106 cells/ml concentration) was harvested in the end of log phase and
comparison with other applied flocculating then used for testing flocculation efficiency.
microorganisms (such as bacteria or fungi) is that
it does not require different cultivation conditions
and therefore avoids additional costs and prevents 2.2. Flocculation experiments
undesired contaminations.
In this study the flocculation efficiency of 2.2.1. Description of the flocculants
commercial PEO compared with PAA and As PAA-based flocculants were used non-ionic Superfloc N-300, anionic
coagulant (i.e., ferric chloride) on harvesting Superfloc A-100, cationic Superfloc C-492 (all three manufactured by
Chlorella vulgaris. The technology of KEMIRA), as PEO-based flocculants – cationic Sibfloc – 718 (manufactured
simultaneously employing coagulant and by Environmental Chemistry research and production company, Russian
flocculant for drinking water purification and Federation), and as inorganic coagulant – FeCl36H2O (Komponent-Reaktiv
sewage treatment was used for harvesting of company, Russian Federation).
microalgae. The flocculated biomass was used as
ballast agent for harvesting of microalgae (Fig. 1).
Also the supernatant after flocculated biomass 2.2.2. Experimental design
separation was reused for microalgae cultivation. The flocculation experiments on C. vulgaris culture broth were carried out
in 250 ml beakers. The flocculant, flocculants mixture or flocculated biomass
were added in varied concentrations. After the addition, the test sample was
 K.V. Gorin et al. / Bioresource Technology 193 (2015) 178–184 181

Fig. 3. Effect of the coagulant and various flocculants mixture adjustment, (a) FeCl36H2O, (b) FeCl36H2O + Superfloc C-492, FeCl36H2O + Sibfloc-718, Superfloc C492 + Sibfloc-718, (c) FeCl36H2O
+ Sibfloc-718 at different sedimental time on the flocculation efficiency of Chlorella vulgaris GKV1.
stirred at 200 rpm during 1 min and then left for 2.2.2.4. Flocculated biomass experiments with flocculant mixtures. The
sedimentation at room temperature. Each coagulants and flocculants were added in the following concentrations to
experiment had a control without flocculant. produce the flocculated biomass: ferric chloride – 100 mg/l + Sibfloc-718–15
All experiments were performed in triplicate. mg/l, Superfloc C-492 25 mg/l, Sibfloc-718 15 mg/l, ferric chloride 50 mg/l
Experiments of flocculation were carrying + Sibfloc-718 7.5 mg/l. The material was stirred at 200 rpm during 1 min
out for 120 min according to another research before it was left to settle for 5 min at room temperature, and then stirred again
(Chen et al., 2013). The special attention was and added to make 5%, 10%, 15% by volume of the native biomass.
focused on flocculation time from 5 to 20 min
as a most appropriate for industrial application. 2.2.3. Flocculation efficiency evaluation
At certain intervals an aliquot was collected at half-height of the beaker and
2.2.2.1. Experiments with individual optical density (OD) was measured using Jenway 6300 spectrophotometer at 680
flocculant. The testing concentrations of nm wavelength. The flocculation efficiency was calculated by equation (Kim et
individual flocculants were: for Superfloc N- al., 2011; Wu et al., 2012): flocculation efficiency (%) = (1 A/B) 100, where A
300 and Superfloc A-100 – 0.2, 0.4, 0.6, 0.8, is OD microalgal suspension after addition of flocculant, and B is OD control
and 1.0 mg/l, for Superfloc C-492 and Sibfloc (without flocculant) at sedimentation time.
– 718 – 5, 15, 25, 35 and 45 mg/l, and for
FeCl36H2O 100, 200, 300, 400 and 500 mg/l.
The same concentration of coagulant and 2.2.4. Reuse of remained culture media after flocculation for
flocculants has been used in paper (Chen et al., cultivation
2013). After flocculation experiment in 250 ml beaker with mixture of ferric chloride
50 mg/l and Sibfloc-718 7.5 mg/l culture broth was centrifuged at 8000 rpm 10
2.2.2.2. Experiments with flocculants mixture. min (Awel MF 20). The supernatant was divide from sludge and 1/10 of Basal
The flocculants mixtures consisted of: ferric medium (Yeh and Chang, 2012) was added. This culture media has been reused
chloride (100; 50; 25; 12.5; 6.25 mg/l) with for C. vulgaris cultivation under conditions described in point 2.1.
Sibfloc-718 (15; 7.5; 4; 2; 1 mg/l), respectively.
3. Results and discussion
2.2.2.3. Flocculated biomass experiments with
individual agents. The coagulants and
flocculants were added in the following 3.1. Effect of different types and dosages of flocculants on flocculation
efficiency of C. vulgaris
concentrations to produce the flocculated
biomass: ferric chloride – 400 mg/l, Superfloc
C-492 – 25 mg/l and Sibfloc – 718 – 15 mg/l.
The material was stirred at 200 rpm during 1
min before it was left to settle for 5 min at room
temperature, and then stirred again and added
to make 5%, 10%, 15% by volume of the native
biomass.
182 K.V. Gorin et al. / Bioresource Technology 193 (2015) 178–184

In early stage of the study, different flocculants achieved in 120 min for both flocculants. However, note that the efficiency, over

Fig. 4. Effect of the flocculated biomass adjustment (by the volume), (a) 5%, (b) 10%, (c) 15%, (d) mixture FeCl36H2O + Sibfloc-718, (e) mixture of flocculant and flocculated biomass at different
sedimental time on the flocculation efficiency of Chlorella vulgaris GKV1.
were tested individually. It was the goal of that 5 min of sedimentation, for Sibfloc-718 of 15 mg/l concentration (Fig. 2d) was
stage to pick out the more efficient flocculants for 50%, whereas in Superfloc C-492 case at 25 mg/l concentration (Fig. 2c) it was
further experimentation using coagulant and 40%.
flocculant mixtures, and with flocculated biomass This is in agreement with the published information that non-ionic and
used as ballast agent. From the experiments, it was anionic flocculants are not suitable to flocculate the microalgae C. vulgaris,
found for non-ionic Superfloc N-300 (Fig. 2a) and Chlorella fusca, Scenedesmus subspicatus b Scenedesmus sp., which was
anionic Superfloc A-100 (Fig. 2b) flocculants obtained in (Granados et al., 2012). Here, it is reported that polymer-based
used, that the flocculation efficiency both was of cationic flocculants proved successful in microalgae harvest. Also, as shown
them close to the control experiment. Our study in (König et al., 2014), a weak-charged cationic flocculant FLOPAM FO 4240
has shown that the PEO-based cationic flocculant SH was most efficient (over 90%) at 2 mg/l and 4 mg/l concentrations for
Sibfloc-718 has similar efficiency to that of the Conticribra weissflogii microalgae. The successful use of cationic PAA
PAA cationic flocculant Superfloc C-492. Thus, flocculants versus non-ionic and anionic agents was also shown for
sedimentation efficiency of about 80% was Phaeodactylum tricornutum and Neochloris oleoabundans microalgae. With

10 mg/l addition of Zetag 7557 and Synthofloc
5080H flocculants, the flocculation efficiency
for P. tricornutum was 98% and 94%, and 52%
and 36% for N. oleoabundan, respectively (‘t
Lam et al., 2014).
Consequently, a smaller quantity of PEO-
based flocculant is required for efficient
sedimentation of C. vulgaris. This also
indicates that PEO-based flocculants can be
more efficient for C. vulgaris sedimentation.
3.2. Effect of coagulation/flocculation on
flocculation efficiency of C. vulgaris
For the coagulating agent, i.e., ferric
chloride, the highest efficiency over 5 min of
sedimentation was more than 60% achieved at
500 mg/l concentration (Fig. 3a). However,
note that this is a very high concentration, so it
may have unfavorable effects on subsequent
biomass processing. Further increase of the
above-given coagulating agent concentration
did not result in higher flocculation efficiency.
Following the use of coagulant and the
flocculating agents based on different
polymers, the study looked at combined
application of coagulants and flocculants.
Thus, over 5 min of sedimentation the
flocculation efficiency was, for combined use
of ferric chloride with Superfloc C-492 and
ferric chloride with Sibfloc-718, equal to 60%
and 87%, respectively (Fig. 3b). These indicate
that flocculation efficiency of mixtures is much
higher than for these flocculants used
individually. Note also, that the flocculation
efficiency with mixture exceeded 80% in less
than 5 min, which is much faster than with pure
flocculants. Thus, one can conclude, that the
mixed use of ferric chloride and polymeric
cationic flocculant can significantly add to the
flocculation efficiency of C. vulgaris. The
increased flocculation efficiency with
combined use of the two agents is attributed to
differences in their mechanisms, that ferric
chloride is to destabilize the system, so making
the cells stick between each other; while
polymer (PAA or PEO) is to help the resulting
flocs become larger, allowing their aggregation
with each other. Once the ferric chloride
concentration was taken equal to 100 mg/l, the
biomass harvested has a less concentration of
Fe than in the case where FeCl3, was solely
used, whose maximum of efficiency was
achieved at 500 mg/l concentration. By the
reason of such effective action mixture of
coagulant and flocculant it was interesting
reduce the dosage for lower costs. Fig. 3c
shows the dependence of the flocculation
efficiency from the mixing ratio of the
coagulant and flocculant. It found that the
reduction of iron concentration to 50 mg/l and
flocculant to 7.5 mg/l does not affect on
flocculation efficiency (approximately 90%
within 5 min of sedimentation), while further
diminishing of concentrations reduce
flocculation efficiency of C. vulgaris.
K.V. Gorin et al. / Bioresource Technology 193 (2015) 178–184 183
From Figs. 2c,d and 3b, efficiency of mixture of 15 mg/l Sibfloc-718 and
25 mg/l Superfloc C-492, 25 mg/l Superfloc C-492 and 15 mg/l Sibfloc-718
equal to 25%, 40% and 50% respectively. This means that combined use of
the two cationic flocculants did not show any benefits as compared with them
used individually. Since the two flocculants offer similar mechanisms, an
effect was obtained in this case, which was similar to what was observed with
their concentrations increased to 45 mg/l, when used separately (Fig. 2c and
d). Thus, the experimental results show the use of two flocculants similar in
mechanism is not effective for efficient sedimentation of C. vulgaris.
3.3. Effect of ballast agent on flocculation efficiency of C. vulgaris
This study looked into the possibility to add the already flocculated
biomass into microalgal culture broth. Thus, the biomasses added were both
flocculated by different flocculants, and those flocculated by coagulant and
flocculant mixtures. In one case, it was biomass flocculated by agents that
showed their efficiency, in particular, Superfloc C-492, Sibfloc-718 and
FeCl3. Also, the
Fig. 5. The cultivation of Chlorella vulgaris GKV1 on reused grow medium.
experiments used the mixture that also proved to be the most efficient earlier, i.e.,
Sibfloc-718 and FeCl3.
The highest efficiency was achieved using biomass flocculated by Sibfloc-
718 and FeCl3 mixture. Note, that the sedimentation efficiency with its addition
would reach about 50% and keep at this level for quite a time (Fig. 4a–d). Thus,
this method is less effective than the case with Sibfloc-718 and FeCl3 mixture.
More interesting results were obtained by joint use of flocculant and the
flocculated biomass as a precipitating agent (Fig. 4e). After 5 min of
sedimentation the maximum efficiency over 90% was achieved by using mixture
of coagulant – ferric chloride and Sibfloc-718 with flocculated biomass, slight
lower 85% – for mixture Sbifloc-718 with flocculated biomass, whereas the
mixture Superfloc C-492 with flocculated biomass did not posses such efficiency.
3.4. Experiments with remained culture media after flocculation
Reusing of supernatant after separation flocculated biomass microalgae can
substantially reduce influence on environment and decrease water loss. This
possibility has been demonstrated in several works (Kim et al., 2011; Wu et al.,
2012). In our study, we also investigated this possibility reusing of supernatant
(Fig. 5). It has been found that algae C. vulgaris was grown on reusable medium
as well as on fresh medium. Thus, supernatant after separation flocculated
biomass can be reused for cultivation microalgae.
4. Conclusions
The data given above bring to the conclusion, that the PEO Sibfloc-718
together with PAA cationic flocculants can be efficiently used for harvesting
184 K.V. Gorin et al. / Bioresource Technology 193 (2015) 178–184
biomass of C. vulgaris GKV1. Sedimentation
efficiency of about 80% was achieved in 120 min
for both flocculants. Combined use of ferric
chloride and polymeric cationic flocculant is
shown to allow significant increase in biomass
sedimentation rate up to 80% in the first five
minutes. Furthermore, partial addition of
flocculated biomass could serve as a ballast agent,
where the flocculation efficiency would be up to
90% in the first 5 min. In this case we can speak
about reducing the dosage of flocculant, at least,
two times.
Acknowledgements
This work was made with financial support of
the Ministry of Education and Science of the
Russian Federation. The number of agreement is
No. 14.607.21.0034. Unique identification number
of applied research is RFMEFI60714X0034.
References
Aguilar, M.I., Sáez, J., Lloréns, M., Soler, A., Ortuño, J.F.,
Meseguer, V., Fuentes, A., 2005. Improvement of
coagulation–flocculation process using anionic
polyacrylamide as coagulant aid. Chemosphere 58, 47–
56.
Banerjee, C., Gupta, P., Mishra, S., Sen, G., Shukla, P.,
Bandopadhyay, R., 2012. Study of polyacrylamide
grafted starch based algal flocculation towards
applications in algal biomass harvesting. Int. J. Biol.
Macromol. 51, 456461.
Ben-Amotz, A., Avron, M., 1990. The biotechnology of
cultivating the halotolerant alga Dunaliella. Trends
Biotechnol. 8, 121125.
Bratby, J., 2006. Coagulation and Flocculation in Water and
Wastewater Treatment, second ed. IWA Publishing,
London, Seattle (pp. 450).
Chang, Y.-R., Lee, D.-J., 2012. Coagulation–membrane
filtration of Chlorella vulgaris at different growth phases.
Drying Technol. 30, 13171322.
Chen, L., Wang, C., Wang, W., Wei, J., 2013. Optimal
conditions of different flocculation methods for
harvesting Scenedesmus sp. cultivated in an open-pond
system. Biores. Technol. 133, 915.
El-Diwani, G.I., El-Rafei, S.A., Hawash, S.I., El-Enin, S.A.,
2011. Optimized flocculation of microalgae for fuel oil
and antioxidant production. Der Chem. Sin. 2, 1225.
Granados, M.R., Acién, F.G., Gómez, C., Fernández-Sevilla,
J.M., Grima, E.M., 2012. Evaluation of flocculants for the
recovery of freshwater microalgae. Bioresour. Technol.
118, 102110.
Grima, E.M., Belarbi, E.-H., Fernández, F.G.A., Medina,
A.R., Chisti, Y., 2003. Recovery of microalgal biomass
and metabolites: process options and economics.
Biotechnol. Adv. 20, 491515.
Harith, Z.T., Yusoff, F.M., Mohamed, M.S., Din, M.S.M.,
Ariff, A.B., 2009. Effect of different flocculants on the
flocculation performance of microalgae, Chaetoceros
Calcitrans, cells. Afr. J. Biotechnol. 8, 59715978.
Haveren, J., Scott, E.L., Sanders, J., 2008. Bulk chemicals
from biomass. Biofuel. Bioprod. Biorg. 2, 4157.
Jiang, J.Q., 2015. The role of coagulation in water treatment.
Curr. Opin. Chem. Eng. 8, 3644.
Jiménez, B., Chávez, A., Hernández, C., 1999. Alternative
treatment for wastewater destined for agricultural use.
Water Sci. Technol. 40, 355362.
Kim, D.G., La, H.J., Ahn, C.Y., Park, Y.H., Oh, H.M., 2011.
Harvest of Scenedesmus sp with bioflocculant and reuse
of culture medium for subsequent highdensity cultures.
Biores. Technol. 102, 31633168.
König, R.B., Sales, R., Roselet, F., Abreu, P.C., 2014.
Harvesting of the marine microalga Conticribra
weissflogii (Bacillariophyceae) by cationic polymeric
flocculants. Biom. Bioeng. 68, 16.
Lee, S.J., Kim, S.-B., Kim, J.-E., Kwon, G.-S., Yoon, B.-D., Oh, H.-M., 1998. Effects of harvesting
method and growth stage on the flocculation of the green alga Botryococcus braunii. Lett. Appl.
Microbiol. 27, 1418.
Lee, K.E., Teng, T.T., Morad, N., Poh, B.T., Mahalingam, M., 2011. Flocculation activity of novel
ferric chloride-polyacrylamide (FeCl3-PAM) hybrid polymer.
Desalination 266, 108113.
López-Maldonado, E.A., Oropeza-Guzman, M.T., Jurado-Baizaval, J.L., Ochoa-Terán, A.,
2014. Coagulation–flocculation mechanisms in wastewater treatment plants through zeta
potential measurements. J. Hazard. Mater. 279, 110.
Rwehumbiza, V.M., Harrison, R., Thomsen, L., 2012. Alum-induced flocculation of
preconcentrated Nannochloropsis salina: residual aluminium in the biomass, FAMEs and
its effects on microalgae growth upon media recycling. Chem. Eng. J. 200202, 168175.
Salim, S., Bosma, R., Vermuë, M.H., Wijffels, R.H., 2011. Harvesting of microalgae by bio-
flocculation. J. Appl. Phycol. 23, 849855.
Schlesinger, A., Eisenstadt, D., Bar-Gil, A., Carmely, H., Einbinder, S., Gressel, J., 2012.
Inexpensive non-toxic flocculation of microalgae contradicts theories; overcoming a major
hurdle to bulk algal production. Biotechnol. Adv. 30, 10231030.
Seybold, C.A., 1994. Polyacrylamide review: soil conditioning and environmental fate. Commun.
Soil Sci. Plant Anal. 25, 2171–2185.
Surendhiran, D., Vijay, M., 2013. Influence of bioflocculation parameters on harvesting
Chlorella salina and its optimization using response surface methodology. J. Environ.
Chem. Eng. 1, 10511056.
‘t Lam, G.P., Vermuë, M.H., Olivieri, G., van den Broek, L.A.M., Barbosa, M.J., Eppink,
M.H.M., Wijffels, R.H., Kleinegris, D.M.M., 2014. Cationic polymers for successful
flocculation of marine microalgae. Biores. Technol. 169, 804807.
Talukder, M.M.R., Das, P., Wu, J.C., 2014. Immobilization of microalgae on exogenous fungal
mycelium: a promising separation method to harvest both marine and fresh water
microalgae. Biochem. Eng. J. 91, 5357.
Thompson, M.S., Vadala, T.P., Vadala, M.L., Lin, Y., Riffle, J.S., 2008. Synthesis and
applications of heterobifunctional poly(ethylene oxide) oligomers. Polymer 49, 345–373.
Tripathy, T., De, B.R., 2006. Flocculation: a new way to treat the waste water. J. Phys. Sci. 10,
93127.
Uduman, N., Qi, Y., Danquah, M.K., Forde, G.M., Hoadley, A., 2010. Dewatering of microalgal
cultures: a major bottleneck to algae-based fuels. J. Renew. Sust. Energy 2, 012701.
Vandamme, D., Foubert, I., Muylaert, K., 2013. Flocculation as a low-cost method for
harvesting microalgae for bulk biomass production. Trend. Biotechnol. 31, 233239.
Vereschagin, E.I., Han, D.H., Troitsky, A.W., Grishin, O.V., Petrov, S.E., Gulyaeva, E.P.,
Bogdanova, L.A., Korobeinikov, M.V., Auslender, V.L., 2001. Radiation technology in
the preparation of polyethylene oxide hydrophilic gels and immobilization of proteases for
use in medical practice. Arch. Pharm. Res. 2001, 229233.
Wan, C., Alam, M.A., Zhao, X.Q., Zhang, X.Y., Guo, S.L., Ho, S.H., Chang, J.S., Bai, F.W.,
2015. Current progress and future prospect of microalgal biomass harvest using various
flocculation technologies. Bioresour. Technol. 184, 251–257.
Wileman, A., Ozkan, A., Berberoglu, H., 2011. Rheological properties of algae slurries for
minimizing harvesting energy requirements in biofuel production. Bioresour. Technol.
104, 432439.
Wu, Z., Zhu, Y., Huang, W., Zhang, C., Li, T., Zhang, Y., Li, A., 2012. Evaluation of
flocculation induced by pH increase for harvesting microalgae and reuse of flocculated
medium. Biores. Technol. 110, 496502.
Yeh, K.-L., Chang, J.-S., 2012. Effects of cultivation conditions and media composition. Biores.
Technol. 105, 120127.
Zhang, J., Hu, B., 2012. A novel method to harvest microalgae via co-culture of filamentous fungi
to form cell pellets. Biores. Technol. 114, 529535.